Gaining an understanding of the mechanisms by which cells process and respond to extracellular cues has become a major goal in many areas of bi- ogy and has attracted the attentions of almost every traditional discipline within the biological sciences. At the heart of these divergent endeavors are common methods that can aid biochemists, physiologists, and pharmacologists in ta- ling the specific questions addressed by their research. In Receptor Signal Transduction Protocols, a diverse array of meth- ologies employed to interrogate ligand--receptor and receptor-effector int- actions are described by authors who have devised and successfully applied them. The authors blend excellent descriptions and applications of fairly well established methodologies with new technologies at the cutting-edge of signal transduction research and as such I hope the present volume will complement and extend a previous excellent volume in this series edited by David Kendall and Stephen Hill (Methods Molecular Biology, vol. 41, Signal Transd- tion Protocols).

Klappentext

An outstanding collection of state-of-the-art methods from leading experts for interrogating ligand-receptor and receptor-effector interactions. The methods emphasize the regulation of G protein-coupled receptors and focus on receptor characterization, methods for investigating how receptors couple to G proteins, and approaches to studying how receptor expression and function is regulated. The easily reproducible protocols include chimeric receptor construction, antibody generation, transient and stable transfection methods, in situ hybridization, as well as immunocytochemistry, functional assays of G protein function, antisense methods for defining receptor-effector coupling pathways, and many more.

Zusammenfassung
Presents state-of-the-art methods for interrogating ligand-receptor and receptor effector interactions. This book should be of interest to neuroscientists, pharmacologists, medicinal chemists, and molecular and cell biologists.

Inhalt
Methods for Characterizing Receptors.- Radioligand-Binding Methods for Membrane Preparations and Intact Cells.- Site-Directed Mutagenesis and Chimeric Receptors in the Study of Receptor-Ligand Binding.- Approaches to the Stable Transfection of G Protein-Coupled Receptors.- Methods for Transient Expression of Hetero-Oligomeric Ligand-Gated Ion Channels.- The Generation of Receptor-Selective Antibodies.- In Situ Hybridization.- lmmunocytochemical Methods for Investigating Receptor Localization.- Methods for Studying Receptor Coupling Mechanisms.- Measurement of Agonist-Stimulated [35S]GTP?S Binding to Cell Membranes.- Autoradiographic Visualization in Brain of Receptor-G Protein Coupling Using [35S]GTP?S Binding.- Agonist-Induced High-Affinity GTP Hydrolysis as an Index of Receptor-Mediated G Protein Activation in Mammalian Brain Membranes.- Use of Random-Saturation Mutagenesis to Study Receptor-G Protein Coupling.- Identification and Quantitation of G Protein ?-Subunits.- Covalent Modification of G Proteins by Affinity Labeling.- Heterologous Expression of Receptors and Signaling Proteins in Adult Mammalian Sympathetic Neurons by Microinjection.- Nuclear Application of Antisense Oligonucleotides by Microinjection and Ballistomagnetic Transfer to Identify G Protein Heterotrimers Activating Phospholipase C.- Use of Antisense-Generating Plasmids to Probe the Function of Signal Transduction Proteins in Primary Neurons.- Methods for Investigating the Regulation of Receptor Function.- Protocols Employed in the Investigation of G Protein-Coupled Receptor Phosphorylation.- Assay of G Protein-Coupled Receptor Kinase Activity by Rhodopsin Phosphorylation.- Radioligand Binding Measurement of Receptor Sequestration in Intact and Permeabilized Cells.- Regulation of ReceptorExpression.